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Spatio-temporal control of gene expression by physico-chemical means: from in vitro photocontrol to smart drug delivery
Final Report Summary - GENEPHYSCHEM (Spatio-temporal control of gene expression by physico-chemical means: from in vitro photocontrol to smart drug delivery)
The general objective of this ERC project was to develop original physico-chemical methods for the control of gene expression and its applications to create new addressable functionalities through a fine control of protein activity. Among possible triggers, light has been identified as a particularly interesting stimulus as it provides excellent spatio-temporal resolution, biocompatibility, and contactless actuation. Following this way, we have developed new molecular tools for reversible photocontrol of DNA compaction, which has been exploited to optically regulate the activity of various encoded or conjugated proteins, including fluorescent proteins and enzymes. We have also created a new class of photosensitive DNA intercalators, allowing photoreversible DNA hybridization/melting at a constant temperature, in a modification-free and sequence-independent manner for the first time. This new concept opens great perspectives for optical control of DNA-based nanotechnologies (e.g. DNA origamis) and biotechnologies (e.g. DNA amplification by PCR). All of these approaches were combined with the bottom-up reconstitution of DNA/protein micro-environments using microfluidic devices and/or well-defined giant liposomes. We have in particular invented a method for one-pot, cell-free reconstitution of membrane proteins in giant liposomes. Interestingly, the functionality of these so-called giant proteoliposomes can be placed under the control of an external stimulus (light, temperature), for instance to control membrane permeability and trigger cargo release. Finally we have discovered that the amphiphilic photosensitive molecules initially developed to control DNA compaction could be adapted to modulate fluid/fluid interfacial energy, thus allowing us to achieve a broad variety of light-driven microfluidic operations.